PathScan? Phospho-Stat3 (Tyr705) Chemiluminescent Sandwich ELISA Kit

• 产品编号：CST-7149C      品牌：CST       原厂货号：7149C
• 产品分类：ELISA及其它免疫分析 > ELISA > ELISA检测试剂盒
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描述：

PathScan® Phospho-Stat3 (Tyr705) Chemiluminescent Sandwich ELISA Kit 检测内源水平在Tyr705位点磷酸化人和鼠的含有Stat3蛋白的细胞。此试剂盒能够检测已经经内部严格测试的物种的蛋白，但是也不排除检测到其它物种中同源性的蛋白。PathScan® Phospho-Stat3 (Tyr705) Chemiluminescent Sandwich ELISA Kit是固相夹心酶联免疫吸收剂检测试剂盒(ELISA)，它能够检测在Tyr705位点磷酸化Stat3并通过化学发光读出结果。比起颜色反应的检测方法，化学发光ELISAs通常具有更为广泛的动力学范围和更高的灵敏度。此化学发光ELISA，在更小容积的酶标板中，但是显示出更强的信号和更高的灵敏度而且所用的样品更少。Stat3抗体包裹到微孔的外表面。然后与细胞裂解液孵育，phospho 和 nonphospho-Stat3蛋白都能够被抗体捕获。经过严格的冲洗后，phospho-Stat3 mouse mAb检测抗体加入用来检测捕捉到的phospho-Stat3(Tyr705)蛋白。抗鼠IgG，HRP连接抗体用于识别检测抗体的结合。化学发光剂加入进行显色反应。光发射的数量按照相对光单位(RLU)来测量，并按比例换算成在Tyr705位点磷酸化Stat3蛋白的量。Stat3 转录因子是细胞因子和生长因子受体信号通路中重要的信号分子(1)，并且对于小鼠胎儿发育是必须的(2)。Stat3 在很多的人类肿瘤中是组成性激活的(3,4)并有潜在的支配原发性肿瘤的可能(5) ，而且也具有抗凋亡活性(3)。Stat3通过Tyr705位点的磷酸化而激活，从而导致它的二聚化，入核后与DNA结合(6,7)。转录活性似乎受到 MAPK 或 mTOR 信号通路介导的Stat3 Ser727位点的磷酸化的调控(8,9)。Stat3不同剪切形式的表达反应了其生物学功能，Stat3α (86 kDa) 和 Stat3β (79 kDa) 形式的表达比例依赖于细胞类型、配体方向或者细胞发育的成熟阶段(10)。值得注意的是，Stat3β蛋白在C-末端转录活性区域缺少丝氨酸磷酸化位点(8)。

1. Heim, M.H. (2001) J Recept Signal Transduct Res 19, 75-120.
2. Takeda, K. et al. (1997) Proc Natl Acad Sci U S A 94, 3801-4.
3. Catlett-Falcone, R. et al. (1999) Immunity 10, 105-15.
4. Garcia, R. and Jove, R. (1998) J Biomed Sci 5, 79-85.
5. Bromberg, J.F. et al. (1999) Cell 98, 295-303.
6. Darnell, J.E. et al. (1994) Science 264, 1415-21.
7. Ihle, J.N. (1995) Nature 377, 591-4.
8. Wen, Z. et al. (1995) Cell 82, 241-50.
9. Yokogami, K. et al. (2000) Curr Biol 10, 47-50.
10. Biethahn, S. et al. (1999) Exp Hematol 27, 885-94.

原厂资料：

Specificity / Sensitivity

PathScan® Phospho-Stat3 (Tyr705) Chemiluminescent Sandwich ELISA Kit detects endogenous levels of phospho-Stat3 (Tyr705) in human and mouse cells. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

Description

The PathScan® Phospho-Stat3 (Tyr705) Chemiluminescent Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of phospho-Stat3 (Tyr705) protein with a chemiluminescent readout. Chemiluminescent ELISAs often have a wider dynamic range and higher sensitivity than conventional chromogenic detection. This chemiluminescent ELISA, which is offered in low volume microplates, shows increased signal and sensitivity while using smaller sample volume. A Stat3 antibody has been coated on the microwells. After incubation with cell lysates, phospho and nonphospho-Stat3 proteins are captured by the coated antibody. Following extensive washing, a phospho-Stat3 mouse mAb is added to detect the captured phospho-Stat3 (Tyr705) protein. Anti-mouse IgG, HRP-linked antibody is then used to recognize the bound detection antibody. Chemiluminescent reagent is added for signal development. The magnitude of light emission, measured in relative light units (RLU), is proportional to the quantity of phospho-Stat3 (Tyr705) protein.

Background

The Stat3 transcription factor is an important signaling molecule for many cytokines and growth factor receptors (1) and is required for murine fetal development (2). Stat3 is constitutively activated in a number of human tumors (3,4) and possesses oncogenic potential (5) and anti-apoptotic activities (3). Stat3 is activated by phosphorylation at Tyr705, which induces dimerization, nuclear translocation, and DNA binding (6,7). Transcriptional activation seems to be regulated by phosphorylation at Ser727 through the MAPK or mTOR pathways (8,9). Stat3 isoform expression appears to reflect biological function as the relative expression levels of Stat3α (86 kDa) and Stat3β (79 kDa) depend on cell type, ligand exposure, or cell maturation stage (10). It is notable that Stat3β lacks the serine phosphorylation site within the carboxy-terminal transcriptional activation domain (8).

1. Heim, M.H. (2001) J Recept Signal Transduct Res 19, 75-120.
2. Takeda, K. et al. (1997) Proc Natl Acad Sci U S A 94, 3801-4.
3. Catlett-Falcone, R. et al. (1999) Immunity 10, 105-15.
4. Garcia, R. and Jove, R. (1998) J Biomed Sci 5, 79-85.
5. Bromberg, J.F. et al. (1999) Cell 98, 295-303.
6. Darnell, J.E. et al. (1994) Science 264, 1415-21.
7. Ihle, J.N. (1995) Nature 377, 591-4.
8. Wen, Z. et al. (1995) Cell 82, 241-50.
9. Yokogami, K. et al. (2000) Curr Biol 10, 47-50.
10. Biethahn, S. et al. (1999) Exp Hematol 27, 885-94.

注意事项：

Low volume microplate * 12 8-well modules -each module is designed to break apart for 8 tests.
**Kit should be stored at 4°C with the exception of 10X Cell Lysis Buffer, which is stored at –20°C (packaged separately).